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Temperature-dependent growth of protein crystals with temperature-independent solubility: case study of apoferritin
CrystEngComm ( IF 2.6 ) Pub Date : 2020-05-25 , DOI: 10.1039/d0ce00654h Ivaylo L. Dimitrov 1, 2, 3, 4
CrystEngComm ( IF 2.6 ) Pub Date : 2020-05-25 , DOI: 10.1039/d0ce00654h Ivaylo L. Dimitrov 1, 2, 3, 4
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The crystal growth of apoferritin, a protein with negligible temperature dependence of solubility, was investigated in the temperature range of 10–60 °C. An approach which accounts simultaneously for both the interface control and the diffusion limitation of crystal growth in solutions was applied. These two contributions were separately and successfully determined from the experimental data obtained with time-lapse microscopy. It was found that the effective diffusion coefficient of the apoferritin at different temperatures is at least 1.5 times smaller than that predicted by the Stokes–Einstein equation. Analysis of the contribution of interface control revealed that the apoferritin crystal growth proceeded via a nucleation-mediated mechanism rather than in spiral or normal growth mode. The temperature dependence of the size of the 2D nucleus and the work for its formation were determined. The obtained kinetic coefficient indicated that the diffusion-limited growth could become pronounced even for crystals of sizes as small as 20 μm. It also predicted a loss of polyhedral stability at a larger crystal size, although it was not observed experimentally. Finally, a comparative analysis was performed with respect to the time for crystals to grow to a detectable crystal size. This suggested that the lowest crystallization temperature could favor time-consuming processes of polymorph selection.
中文翻译：
具有温度无关溶解度的蛋白质晶体的温度依赖性生长：载铁蛋白的案例研究
在10–60°C的温度范围内研究了载铁蛋白的晶体生长，该蛋白的溶解度对温度的影响可忽略不计。应用了一种同时解决界面控制和溶液中晶体生长扩散限制的方法。这两个贡献是从延时显微镜获得的实验数据中分别成功确定的。发现在不同温度下，载铁蛋白的有效扩散系数至少比斯托克斯-爱因斯坦方程所预测的有效扩散系数小1.5倍。对界面控制作用的分析表明，载铁蛋白晶体的生长通过是成核介导的机制，而不是以螺旋或正常生长方式。确定了二维核尺寸的温度依赖性及其形成功。所获得的动力学系数表明，即使对于小至20μm的晶体，扩散限制的生长也可能变得明显。尽管在实验中没有观察到，但它也预测了在较大晶体尺寸下多面体稳定性的损失。最后，对晶体生长到可检测的晶体尺寸的时间进行了比较分析。这表明最低的结晶温度可能有利于耗时的多晶型物选择过程。
更新日期：2020-07-06
中文翻译：
具有温度无关溶解度的蛋白质晶体的温度依赖性生长：载铁蛋白的案例研究
在10–60°C的温度范围内研究了载铁蛋白的晶体生长，该蛋白的溶解度对温度的影响可忽略不计。应用了一种同时解决界面控制和溶液中晶体生长扩散限制的方法。这两个贡献是从延时显微镜获得的实验数据中分别成功确定的。发现在不同温度下，载铁蛋白的有效扩散系数至少比斯托克斯-爱因斯坦方程所预测的有效扩散系数小1.5倍。对界面控制作用的分析表明，载铁蛋白晶体的生长通过是成核介导的机制，而不是以螺旋或正常生长方式。确定了二维核尺寸的温度依赖性及其形成功。所获得的动力学系数表明，即使对于小至20μm的晶体，扩散限制的生长也可能变得明显。尽管在实验中没有观察到，但它也预测了在较大晶体尺寸下多面体稳定性的损失。最后，对晶体生长到可检测的晶体尺寸的时间进行了比较分析。这表明最低的结晶温度可能有利于耗时的多晶型物选择过程。
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